Aldh2 dependent formaldehyde metabolism fuels purine demands in melanocyte stem cells

In the McSC lineage in zebrafish, Aldh2 activity is an important source of formate through its metabolism of its substrate formaldehyde. This formate is required as input into the folate cycle, in particular de novo purine biosynthesis. This scRNA-seq experiment was conducted to investigate the global effects of Aldh2 inhibition on the transcriptomes of isolated McSCs. Overall design: Tg(mitfa:GFP;crestin:mCherry) zebrafish embryos express GFP and mCherry in regenerating McSCs. Embryos were treated with an Aldh2 inhibitor (CVT-10216), or DMSO (vehicle control) from 24hpf onwards. Embryonic melanocytes were ablated by co-treating with MoTP from 24hpf, and McSC melanocyte regeneration stimulated by washing MoTP out at 64hpf. After 8 hours of melanocyte regeneration, the trunk region of embryos was dissected from the head, and crestin:mCherry+, mitfa:GFP+ or double+ cells isolated by FACs. These cells were then processed for 10x